Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Institute for Stem Cell Biology & Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Biomedicine, Aarhus University, 8000 Aarhus, Denmark; Aarhus Institute of Advanced Studies (AIAS), Aarhus University, 8000 Aarhus, Denmark.
Cell Stem Cell. 2024 Apr 4;31(4):499-518.e6. doi: 10.1016/j.stem.2024.03.002.
Allogeneic hematopoietic stem and progenitor cell transplant (HSCT) of CCR5 null (CCR5Δ32) cells can be curative for HIV-1-infected patients. However, because allogeneic HSCT poses significant risk, CCR5Δ32 matched bone marrow donors are rare, and CCR5Δ32 transplant does not confer resistance to the CXCR4-tropic virus, it is not a viable option for most patients. We describe a targeted Cas9/AAV6-based genome editing strategy for autologous HSCT resulting in both CCR5- and CXCR4-tropic HIV-1 resistance. Edited human hematopoietic stem and progenitor cells (HSPCs) maintain multi-lineage repopulation capacity in vivo, and edited primary human T cells potently inhibit infection by both CCR5-tropic and CXCR4-tropic HIV-1. Modification rates facilitated complete loss of CCR5-tropic replication and up to a 2,000-fold decrease in CXCR4-tropic replication without CXCR4 locus disruption. This multi-factor editing strategy in HSPCs could provide a broad approach for autologous HSCT as a functional cure for both CCR5-tropic and CXCR4-tropic HIV-1 infections.
异体造血干细胞和祖细胞移植（HSCT）的 CCR5 缺失（CCR5Δ32）细胞可以治愈 HIV-1 感染患者。然而，由于异体 HSCT 存在重大风险，CCR5Δ32 匹配的骨髓供体很少，并且 CCR5Δ32 移植不能抵抗 CXCR4 嗜性病毒，因此它不是大多数患者的可行选择。我们描述了一种针对自体 HSCT 的靶向 Cas9/AAV6 基因组编辑策略，可导致 CCR5 和 CXCR4 嗜性 HIV-1 耐药。编辑的人造血干细胞和祖细胞（HSPCs）在体内保持多谱系重殖能力，编辑的原代人 T 细胞强烈抑制 CCR5 嗜性和 CXCR4 嗜性 HIV-1 的感染。修饰率可促进 CCR5 嗜性复制完全丧失，并将 CXCR4 嗜性复制降低多达 2000 倍，而不会破坏 CXCR4 基因座。这种在 HSPCs 中的多因素编辑策略可以为自体 HSCT 提供广泛的方法，作为 CCR5 嗜性和 CXCR4 嗜性 HIV-1 感染的功能性治愈。
